Isolation and Biological Characteristics of Bone Marrow Mesenchymal Stem Cells Derived from Wistar Rats

Abstract: This work was aim to establish a simple， efficient and optimum isolation and culture method of bone marrow mesenchymal stem cells (BMSCs) derived from Wistar rat. This method would lay the foundation for the study of its biological characteristics. In this study， BMSCs-derived from Wistar rats were isolated by the method-combined of whole bone marrow adherent culture and density gradient centrifugation under the sterile environment. The cells which were cultured in routine conditions and the cells which were induced respectively by osteogenic fluid and adipogenic fluid were observed by inverted phase contrast microscope， and the characteristics of cell proliferation and differentiation were analysed by counting cell， Western blot， Real-time PCR and flow cytometry.The results showed that the isolated cells presented typical uniform swirling growth state and long spindle-shaped after the third generation， the growth curve was for “S” shape， the positive rate of cell surface antigen markers CD29，CD34， CD44， CD45 and CD90 were 98.21%， 0.78%， 91.62%， 0.93% and 99.27%， respectively. After adipogenic and osteogenic induction， protein levels of PPAR-γ2， LPL， Runx2 and BMP-2 were significantly increased， and large amounts of lipid droplets and calcium nodulesappeared in the cellsof different induction by the Oil Red O and the Alizarin Red staining， respectively. BMSCs-derived from Wistar rat were successfully isolated and culutured by the method of whole bone marrow adherent culture and density gradient centrifugation. The cells-isolated have the potential abilities of osteogenic and adipogenic differentiation and the biological properties of BMSCs.

CSTR: 32200.14.cjcb.2017.03.0010